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Differential immune microenvironmental features of microsatellite-unstable colorectal cancers according to Fusobacterium nucleatum status

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Abstract

It has been suggested that Fusobacterium nucleatum (Fn) may differentially impact tumor immune responses according to microsatellite instability (MSI) status in colorectal cancers (CRCs). We aimed to reveal the detailed relationship between intratumoral Fn and immune microenvironmental features in MSI-high CRCs. A total of 126 MSI-high CRCs were subjected to analyses for intratumoral Fn DNA load using quantitative PCR and for densities of tumor-infiltrating immune cells, including CD3+ T cells, CD4+ T cells, CD8+ T cells, FoxP3+ T cells, CD68+ macrophages, CD163+ macrophages, and CD177+ neutrophils, at invasive margin (IM) and center of tumor (CT) areas using computational image analysis of immunohistochemistry. Based on the Fn load, the 126 MSI-high CRCs were classified into Fn-high, -low, and -negative subgroups. The Fn-high subset of MSI-high CRCs was significantly correlated with larger tumor size and advanced invasion depth (p = 0.017 and p = 0.034, respectively). Compared with the Fn-low/negative subgroup, Fn-high tumors demonstrated significantly lower density of FoxP3+ cells in both IM and CT areas (p = 0.002 and p = 0.003, respectively). Additionally, Fn-high was significantly associated with elevated CD163+ cell to CD68+ cell ratio in only CT areas of MSI-high CRCs (p = 0.028). In conclusion, the Fn-enriched subset of MSI-high CRCs is characterized by increased tumor growth and invasion and distinct immune microenvironmental features, including decreased FoxP3+ T cells throughout the tumor and increased proportion of M2-polarized macrophages in the tumor center. These findings collectively support that Fn may be linked to pro-tumoral immune responses in MSI-high CRCs.

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Data and/or code availability

All data generated or analyzed during this study are included in this published article and its supplementary information files.

Abbreviations

CRC:

Colorectal cancer

CIMP:

CpG island methylator phenotype

CT:

Center of tumor

DFS:

Disease-free survival

FFPE:

Formalin-fixed, paraffin-embedded

Fn :

Fusobacterium nucleatum

ICI:

Immune checkpoint inhibitor

IHC:

Immunohistochemistry

IM:

Invasive margin

MSI:

Microsatellite instability

OS:

Overall survival

qPCR:

Quantitative polymerase chain reaction

Th:

Helper T cell

TIIC:

Tumor-infiltrating immune cell

TIL:

Tumor-infiltrating lymphocyte

TLS:

Tertiary lymphoid structure

TMA:

Tissue microarray

Treg:

Regulatory T cell

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Acknowledgements

This work was supported by the National Research Foundation of Korea funded by the Korea government (Ministry of Science and ICT) under Grants NRF-2016R1C1B2010627 and NRF-2019R1F1A1059535. The biospecimens for this study were partly provided by the Seoul National University Hospital Cancer Tissue Bank (SNUH CTB). All samples derived from the SNUH CTB were obtained with informed consent under institutional review board-approved protocols. We also thank those who have developed and shared QuPath, an open-source software for digital pathology analysis.

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  1. Ji Ae Lee and Seung-Yeon Yoo have contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, Seoul National University Hospital, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul, 03080, Republic of Korea

    Ji Ae Lee, Seung-Yeon Yoo, Hyeon Jeong Oh, Gyeong Hoon Kang & Jung Ho Kim

  2. Laboratory of Epigenetics, Cancer Research Institute, Seoul National University College of Medicine, Seoul, Republic of Korea

    Ji Ae Lee, Seung-Yeon Yoo, Hyeon Jeong Oh, Seorin Jeong, Nam-Yun Cho, Gyeong Hoon Kang & Jung Ho Kim

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Contributions

Conceptualization: JHK; Formal analysis: JAL, SYY, SJ, NYC, JHK; Funding acquisition: JHK; Investigation: JAL, SYY, JHK; Methodology: JAL, SYY, HJO, SJ, NYC, JHK; Project administration: GHK, JHK; Resources: JAL, HJO; Supervision: GHK, JHK; Validation: JAL, SYY, JHK; Visualization: JAL, SYY, JHK; Writing—original draft: JAL, SYY, JHK; Writing—review & editing: GHK, JHK.

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Correspondence to Jung Ho Kim.

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This study was approved by the institutional review board of Seoul National University Hospital (IRB No. 1805-018-944). This study was performed in accordance with the Declaration of Helsinki.

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All patients were provided written consent for the research or molecular diagnostic use of residual tumor tissues obtained during their surgical treatment.

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Lee, J.A., Yoo, SY., Oh, H.J. et al. Differential immune microenvironmental features of microsatellite-unstable colorectal cancers according to Fusobacterium nucleatum status. Cancer Immunol Immunother 70, 47–59 (2021). https://doi.org/10.1007/s00262-020-02657-x

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